Generation and characterisation of scalable and stable human pluripotent stem cell-derived microvascular-like endothelial cells for cardiac applications
: Majid Qasim A., Ghimire Bishwa R., Merkely Bela, Randi Anna M., Harding Sian E., Talman Virpi, Földes Gabor
Publisher: Springer
: 2024
: Angiogenesis
: Angiogenesis
: Angiogenesis
: 27
: 3
: 561
: 582
: 0969-6970
: 1573-7209
DOI: https://doi.org/10.1007/s10456-024-09929-5
: https://doi.org/10.1007/s10456-024-09929-5
: https://research.utu.fi/converis/portal/detail/Publication/421370031
Coronary microvascular disease (CMD) and its progression towards major adverse coronary events pose a significant health challenge. Accurate in vitro investigation of CMD requires a robust cell model that faithfully represents the cells within the cardiac microvasculature. Human pluripotent stem cell-derived endothelial cells (hPSC-ECs) offer great potential; however, they are traditionally derived via differentiation protocols that are not readily scalable and are not specified towards the microvasculature. Here, we report the development and comprehensive characterisation of a scalable 3D protocol enabling the generation of phenotypically stable cardiac hPSC-microvascular-like ECs (hPSC-CMVECs) and cardiac pericyte-like cells. These were derived by growing vascular organoids within 3D stirred tank bioreactors and subjecting the emerging 3D hPSC-ECs to high-concentration VEGF-A treatment (3DV). Not only did this promote phenotypic stability of the 3DV hPSC-ECs; single cell-RNA sequencing (scRNA-seq) revealed the pronounced expression of cardiac endothelial- and microvascular-associated genes. Further, the generated mural cells attained from the vascular organoid exhibited markers characteristic of cardiac pericytes. Thus, we present a suitable cell model for investigating the cardiac microvasculature as well as the endothelial-dependent and -independent mechanisms of CMD. Moreover, owing to their phenotypic stability, cardiac specificity, and high angiogenic potential, the cells described within would also be well suited for cardiac tissue engineering applications.
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The research was funded by the British Heart Foundation (RE/13/4/30184 and RM/17/1/33377), the Medical Research Council neg 10.1007/s10456-024-09929-5 10.1007/s10456-024-09929-5 (MR/R025002/1), the Research Council of Finland (projects 321564; 353109; 328909), the Finnish Foundation for Cardiovascular Research, Sigrid Jus?lius Foundation, and the Higher Education Institutional Excellence Programme of the Ministry of Human Capacities in Hungary, within the framework of the Therapeutic Development thematic programme of the Semmelweis University, the Hungarian National Research, Development and Innovation Fund (RRF-2.3.1-21-2022-00003, TKP2021-EGA-23, NKFIH 146125). Open access funding provided by Semmelweis University.
